LOCUS       pPuroG1__4304_          5694 bp    DNA        circular     19-MAR-2022
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Golden Gate reaction:
COMMENT     pHygG minus hyg PCR from 386 (BsaI) to 362 (BsaI) 
COMMENT     puro PCR1 from 8 (BsaI) to 592 (BsaI) 
COMMENT     Puro PCR2 from 8 (BsaI) to 122 (BsaI) 
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     promoter        2..363
                     /note="Prps-0"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note=""/ApEinfo_revcolor=pink; /ApEinfo_fwdcolor=pink;"
                     /locus_tag="Prps-0"
                     /label="Prps-0"
                     /ApEinfo_label="Prps-0"
                     /ApEinfo_fwdcolor="#339933"
                     /ApEinfo_revcolor="#339933"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             365..948
                     /locus_tag="PuroR"
                     /label="PuroR"
                     /ApEinfo_label="PuroR"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          564..614
                     /locus_tag="SynIntronI"
                     /label="SynIntronI"
                     /ApEinfo_label="SynIntronI"
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="#ffff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          815..865
                     /locus_tag="SynIntronII"
                     /label="SynIntronII"
                     /ApEinfo_label="SynIntronII"
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="#ffff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             949..1063
                     /locus_tag="PuroR(1)"
                     /label="PuroR(1)"
                     /ApEinfo_label="PuroR"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             4003..4662
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(1546..1947)
                     /locus_tag="myo-2 3' UTR"
                     /label="myo-2 3' UTR"
                     /ApEinfo_label="myo-2 3' UTR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2956..2958)
                     /locus_tag="pMYO-2"
                     /label="pMYO-2"
                     /ApEinfo_label="pMYO-2"
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2962..2995)
                     /locus_tag="FRT"
                     /label="FRT"
                     /ApEinfo_label="FRT"
                     /ApEinfo_fwdcolor="#800000"
                     /ApEinfo_revcolor="#008080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2769..2819)
                     /locus_tag="Synthetic Intron 1"
                     /label="Synthetic Intron 1"
                     /ApEinfo_label="Synthetic Intron 1"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(3147..3180)
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2589..2639)
                     /locus_tag="Synthetic Intron 2"
                     /label="Synthetic Intron 2"
                     /ApEinfo_label="Synthetic Intron 2"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      3236..3542
                     /locus_tag="F1 ori"
                     /label="F1 ori"
                     /ApEinfo_label="F1 ori"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(join(2011..2130,2182..2406,2458..2588,
                     2640..2768,2820..2925))
                     /locus_tag="NeonGreen_Cel"
                     /label="NeonGreen_Cel"
                     /ApEinfo_label="NeonGreen_Cel"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="#00ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     transit_peptide complement(1954..2010)
                     /locus_tag="egl-13 NLS"
                     /label="egl-13 NLS"
                     /ApEinfo_label="egl-13 NLS"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     rep_origin      4760..5447
                     /locus_tag="ColE1 origin"
                     /label="ColE1 origin"
                     /ApEinfo_label="ColE1 origin"
                     /ApEinfo_fwdcolor="#808080"
                     /ApEinfo_revcolor="#808080"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(2131..2181)
                     /locus_tag="Synthetic Intron 4"
                     /label="Synthetic Intron 4"
                     /ApEinfo_label="Synthetic Intron 4"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     3'UTR           1068..1419
                     /locus_tag="unc-54 3'UTR"
                     /label="unc-54 3'UTR"
                     /ApEinfo_label="unc-54 3'UTR"
                     /ApEinfo_fwdcolor="#990000"
                     /ApEinfo_revcolor="#cc0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     sig_peptide     complement(2926..2958)
                     /locus_tag="SV40 nls"
                     /label="SV40 nls"
                     /ApEinfo_label="SV40 nls"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="#ff0080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(2407..2457)
                     /locus_tag="Synthetic Intron 3"
                     /label="Synthetic Intron 3"
                     /ApEinfo_label="Synthetic Intron 3"
                     /ApEinfo_fwdcolor="#ff80c0"
                     /ApEinfo_revcolor="#ff80c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    3016..3049
                     /locus_tag="FRT3"
                     /label="FRT3"
                     /ApEinfo_label="FRT3"
                     /ApEinfo_fwdcolor="#804000"
                     /ApEinfo_revcolor="#008040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(3049..3103)
                     /locus_tag="MCS"
                     /label="MCS"
                     /ApEinfo_label="MCS"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 cATTTTTGCT TTCGTCGTAA ATCTACACAC GCGTCTCTTC CGTGCGAGAG TCCAAGCCAG
       61 CAGCCAAATT CGTTGACTGA GTATTCAACG TTTATACGTT GTCGGCAACG AGAAATAGGA
      121 AAATGCATCG GGAAATGTTC TTTTTTCGAT TTTTTCCAAG GTTTTGACAA ATTTTACCAC
      181 GAATTTTGCT ATGTTTTCAA TTAAAAAATA TGTTATTCAA CTGTTTCTAT GAGGAAAATA
      241 AGGCTTTGCA TGTAATTTTC TTATTCAGCA TAATTTTTAA TTAATTTGAA TTTTCTGTCC
      301 TAACGTTTAT TTTGTTTTCT TGGTTATGAC TGATCTGAAA TTAATTTTTG AATTTTAAGG
      361 TAATATGACC GAGTACAAGC CCACGGTGCG CCTCGCCACC CGCGACGACG TCCCCAGGGC
      421 CGTACGCACC CTCGCCGCCG CGTTCGCCGA CTACCCCGCC ACGCGCCACA CCGTCGATCC
      481 GGACCGCCAC ATCGAGCGGG TCACCGAGCT GCAAGAACTC TTCCTCACGC GCGTCGGGCT
      541 CGACATCGGC AAGGTGTGGG TCGgtaagtt taaacatata tatactaact aaccctgatt
      601 atttaaattt tcagCGGACG ACGGCGCCGC GGTGGCGGTC TGGACCACGC CGGAGAGCGT
      661 CGAAGCGGGG GCGGTGTTCG CCGAGATCGG CCCGCGCATG GCCGAGTTGA GCGGTTCCCG
      721 GCTGGCCGCG CAGCAACAGA TGGAAGGCCT CCTGGCGCCG CACCGGCCCA AGGAGCCCGC
      781 GTGGTTCCTG GCCACCGTCG GCGTCTCGCC CGACgtaagt ttaaacatga ttttactaac
      841 taactaatct gatttaaatt ttcagCACCA GGGCAAGGGT CTGGGCAGCG CCGTCGTGCT
      901 CCCCGGAGTG GAGGCGGCCG AGCGCGCCGG GGTGCCCGCC TTCCTGGAAA CCTCCGCGCC
      961 CCGCAACCTC CCCTTCTACG AGCGGCTCGG CTTCACCGTC ACCGCCGACG TCGAGGTGCC
     1021 CGAAGGACCG CGCACCTGGT GCATGACCCG CAAGCCCGGT GCCTGAGTCC AATTACTCTT
     1081 CAACATCCCT ACATGCTCTT TCTCCCTGTG CTCCCACCCC CTATTTTTGT TATTATCAAA
     1141 AAACTTCTCT TAATTTCTTT GTTTTTTAGC TTCTTTTAAG TCACCTCTAA CAATGAAATT
     1201 GTGTAGATTC AAAAATAGAA TTAATTCGTA ATAAAAAGTC GAAAAAAATT GTGCTCCCTC
     1261 CCCCCATTAA TAATAATTCT ATCCCAAAAT CTACACAATG TTCTGTGTAC ACTTCTTATG
     1321 TTTTTTACTT CTGATAAATT TTTTTGAAAC ATCATAGAAA AAACCGCACA CAAAATACCT
     1381 TATCATATGT TACGTTTCAG TTTATGACCG CAATTTTTAG ACCtgttcga ggacaactgg
     1441 gaaactttta caaacttaca acatggttca gacaagttta aaaatttaag tttgagaatc
     1501 aacgggactt tttctttttt cgtttttttt aattgtgtat gctttgaaga ttttcaaatt
     1561 tattctagtt tcaataaaaa cagtaatata attgagtggg agtttttttg ctgtgatgtt
     1621 tttggtgtta tttacagtga tcttcttcta aatagaaaat caatggttta ttgatcaaaa
     1681 atattgagaa gtgtcgaatc aattgaagca ttgaaggaca aatttccact atcctacaaa
     1741 tccgatgcgt aaaaattatt aaacaaagtg aaaaaaatgc atttgaagaa aaaaaacaac
     1801 agagaaggtg ttgatgggct tttattgtga cacgttgaat gtgtatgtgt tactgtgcag
     1861 tggtaggttt gggggacgta ggggaagaga aacgggtaaa gagattgaaa gaaaagagag
     1921 aaacgagatg gaattagagc agttgatcta CTTGGCGAGC TTCTTGGCGT TCTCGGAGAG
     1981 CTTGGTTGGG TTGGCCTTAC GACGACGGGA GGCCTTGTAG AGTTCATCCA TTCCCATCAC
     2041 ATCTGTGAAG GCCTTTTGCC ACTCTTTGAA GTTCAGCTCT GTCTTTGAGT GCTTGAGTTC
     2101 TGTCTTTCGG AACACGTACA TTGGTTGATT ctgaaaattt aaatcagatt agttagttag
     2161 taaaatcatg tttaaactta cCTTGAGGTA GTTGGCGGCC ATTGGCTTAG CGAATGTGTA
     2221 TGTTGTTCGG GCTGTCGATC GGTATCGCTT TCCGTTTCCA GTAGTGTACG ACCACTTGAA
     2281 TGTCGAGATG ATTGTCTTAT CATTTGGGTA TGTCTTTTTC GATCGGCACC AATCAGCAGC
     2341 TGTGAGTGAG TTTGTCATCA CTGGTCCATC AGCTGGGAAT CCTGTTCCCT TAACTTGAGC
     2401 CTCTCCctga aaatttaaat atgtatggtt agttagtacc gaactgttta aacttacCTT
     2461 GATGTGCGAT CCCTCGTATG TGTATCGGTA GTTCACTGTG AGCGAGGCTC CATCCTCGAA
     2521 TTGCATTGTT CGGTGAACTT GGTATCCCGA TCCATCCACC ATGGCAGCTT GGAATGGTGA
     2581 CATTCCATct gaaaatttaa atattactcg ttagttagta gaactcagtt taaacttacC
     2641 TGGGTATGGG AGGTATTGGT GGAATCCGTA TCCGATGTGT GGCACGAGAA TCCATGGCGA
     2701 GAATTGCAGA TCTCCCTTTG TCGACTTGAG GTTGAGTTCC TCGTATCCAT CGTTTGGGTT
     2761 TCCAGTTCct gaaaatttaa ataatcaggg ttagttagta tatatatgtt taaacttacC
     2821 TTGTCCGACC ATATCGAAAT CCACTCCGTT GATCGATCCG AAGATGTGGA GTTCGTGTGT
     2881 AGCTGGGAGT GAAGCCATGT TATCCTCTTC TCCCTTCGAC ACCATCGACA CCTTACGCTT
     2941 CTTCTTTGGG GATCCCATTT TGAAGTTCCT ATACtttcta gaGAATAGGA ACTTCACCga
     3001 tatcgaattc ctgcaGAAGT TCCTATTCtt caaataGTAT AGGAACTTCc cgggTACCGA
     3061 AGAGCatGCa aCGCGTggtt accTGCAGGC TCTTCGCCAC TAGttctaga gcggccgcca
     3121 ccgcggtgga gctccaattc gccCGTATAA CTTCGTATAA TGTATGCTAT ACGAAGTTAT
     3181 CCATCTAGag cggccgccac cgcggtggag ctccgacgac gcgccctgta gcggcgcatt
     3241 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3421 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3541 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3601 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3661 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3781 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3841 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3901 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3961 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4021 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4081 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4141 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4201 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4261 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4321 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4381 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4441 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4501 tctggagccg gtgagcgtgg atctcgcggt atcattgcag cactggggcc agatggtaag
     4561 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4621 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4981 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5521 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5581 gagtcagtga gcgaggaagc ggaattcgcc caatacgcaa accgcctctc cccgcgcgtt
     5641 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggat
//
